LOCUS       sequence-311738-       10360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeted A to G  in rice protoplasts .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10360)
  TITLE     Gao Lab Plasmids
REFERENCE   2  (bases 1 to 10360)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: PABE-7-NG
FEATURES             Location/Qualifiers
     source          1..10360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             7216..7224
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             7316..7336
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7349..7369
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7382..7402
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7460..7636
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(8190..8206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8190..8206)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8203..8225)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8214..8230
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8238..8244,8245..8262,8263..8268))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8283..8304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8421..8438)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8592..9180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8672..8691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9351..10142,10143..10211))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9974..9993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10212..10316)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagctcggta cctgacccgg tcgtgcccct ctctagagat aatgagcatt gcatgtctaa
       61 gttataaaaa attaccacat attttttttg tcacacttgt ttgaagtgca gtttatctat
      121 ctttatacat atatttaaac tttactctac gaataatata atctatagta ctacaataat
      181 atcagtgttt tagagaatca tataaatgaa cagttagaca tggtctaaag gacaattgag
      241 tattttgaca acaggactct acagttttat ctttttagtg tgcatgtgtt ctcctttttt
      301 tttgcaaata gcttcaccta tataatactt catccatttt attagtacat ccatttaggg
      361 tttagggtta atggttttta tagactaatt tttttagtac atctatttta ttctatttta
      421 gcctctaaat taagaaaact aaaactctat tttagttttt ttatttaata atttagatat
      481 aaaatagaat aaaataaagt gactaaaaat taaacaaata ccctttaaga aattaaaaaa
      541 actaaggaaa catttttctt gtttcgagta gataatgcca gcctgttaaa cgccgtcgac
      601 gagtctaacg gacaccaacc agcgaaccag cagcgtcgcg tcgggccaag cgaagcagac
      661 ggcacggcat ctctgtcgct gcctctggac ccctctcgat cgagagttcc gctccaccgt
      721 tggacttgct ccgctgtcgg catccagaaa ttgcgtggcg gagcggcaga cgtgagccgg
      781 cacggcaggc ggcctcctcc tcctctcacg gcaccggcag ctacggggga ttcctttccc
      841 accgctcctt cgctttccct tcctcgcccg ccgtaataaa tagacacccc ctccacaccc
      901 tctttcccca acctcgtgtt gttcggagcg cacacacaca caaccagatc tcccccaaat
      961 ccacccgtcg gcacctccgc ttcaaggtac gccgctcgtc ctcccccccc ccccctctct
     1021 accttctcta gatcggcgtt ccggtccatg gttagggccc ggtagttcta cttctgttca
     1081 tgtttgtgtt agatccgtgt ttgtgttaga tccgtgctgc tagcgttcgt acacggatgc
     1141 gacctgtacg tcagacacgt tctgattgct aacttgccag tgtttctctt tggggaatcc
     1201 tgggatggct ctagccgttc cgcagacggg atcgatttca tgattttttt tgtttcgttg
     1261 catagggttt ggtttgccct tttcctttat ttcaatatat gccgtgcact tgtttgtcgg
     1321 gtcatctttt catgcttttt tttgtcttgg ttgtgatgat gtggtctggt tgggcggtcg
     1381 ttctagatcg gagtagaatt aattctgttt caaactacct ggtggattta ttaattttgg
     1441 atctgtatgt gtgtgccata catattcata gttacgaatt gaagatgatg gatggaaata
     1501 tcgatctagg ataggtatac atgttgatgc gggttttact gatgcatata cagagatgct
     1561 ttttgttcgc ttggttgtga tgatgtggtg tggttgggcg gtcgttcatt cgttctagat
     1621 cggagtagaa tactgtttca aactacctgg tgtatttatt aattttggaa ctgtatgtgt
     1681 gtgtcataca tcttcatagt tacgagttta agatggatgg aaatatcgat ctaggatagg
     1741 tatacatgtt gatgtgggtt ttactgatgc atatacatga tggcatatgc agcatctatt
     1801 catatgctct aaccttgagt acctatctat tataataaac aagtatgttt tataattatt
     1861 ttgatcttga tatacttgga tgatggcata tgcagcagct atatgtggat ttttttagcc
     1921 ctgccttcat acgctattta tttgcttggt actgtttctt ttgtcgatgc tcaccctgtt
     1981 gtttggtgtt acttctgcaa agcttatgtc cgaggtggag ttctctcacg agtattggat
     2041 gaggcacgct cttacacttg ctaagagagc ttgggacgaa agagaagtgc cagttggcgc
     2101 cgttcttgtg cataataata gggtgatcgg cgagggttgg aatagaccaa ttggaaggca
     2161 tgatccaaca gctcacgcag agattatggc tctcagacaa ggcggcctcg ttatgcagaa
     2221 ctacaggctc attgacgcta cactctacgt gacactcgaa ccttgcgtta tgtgcgccgg
     2281 agctatgatt cattctagga ttggcagggt cgtgtttgga gctagggacg ctaaaacagg
     2341 agccgccgga tctcttatgg acgtgttgca tcatccaggc atgaaccata gggtggagat
     2401 tacagagggc attcttgcag acgagtgcgc tgctcttctt tccgatttct tcaggatgag
     2461 aaggcaggag attaaggccc agaagaaggc tcagtcttct acagatagcg gaggatcttc
     2521 cggaggatct agcggctccg agacaccagg aacatccgaa agcgctacac cagaatctag
     2581 cggaggctct tccggaggat cttctgaagt ggagttctcc cacgagtatt ggatgaggca
     2641 cgctcttaca cttgctaaaa gggctaggga cgaaagggaa gttccagttg gagctgttct
     2701 cgtgctcaat aacagggtga ttggcgaggg ttggaataga gccattggac tccatgatcc
     2761 aacagctcac gcagagatta tggctcttag acaaggcggc ctcgttatgc agaattacag
     2821 actcatcgac gccacactct acgttacctt cgaaccttgc gttatgtgcg ccggagctat
     2881 gatccattct aggattggca gggtcgtgtt cggcgttaga aacgctaaga caggagctgc
     2941 aggctctctt atggacgttc ttcattaccc aggcatgaat catagagtgg agatcacaga
     3001 aggcattctt gcagacgagt gcgcagctct cctttgctat ttcttcagga tgccgaggca
     3061 agttttcaac gctcagaaga aggcccagtc ttctacagat tccggcggat cttctggagg
     3121 atctagcggc tccgagacac caggaacatc cgaatccgct acaccagagt cttctggagg
     3181 atctagcgga ggatctctta aggacaagaa gtactcgatc ggcctcgcca ttgggactaa
     3241 ctctgttggc tgggccgtga tcaccgacga gtacaaggtg ccctcaaaga agttcaaggt
     3301 cctgggcaac accgatcggc attccatcaa gaagaatctc attggcgctc tcctgttcga
     3361 cagcggcgag acggctgagg ctacgcggct caagcgcacc gcccgcaggc ggtacacgcg
     3421 caggaagaat cgcatctgct acctgcagga gattttctcc aacgagatgg cgaaggttga
     3481 cgattctttc ttccacaggc tggaggagtc attcctcgtg gaggaggata agaagcacga
     3541 gcggcatcca atcttcggca acattgtcga cgaggttgcc taccacgaga agtaccctac
     3601 gatctaccat ctgcggaaga agctcgtgga ctccacagat aaggcggacc tccgcctgat
     3661 ctacctcgct ctggcccaca tgattaagtt caggggccat ttcctgatcg agggggatct
     3721 caacccggac aatagcgatg ttgacaagct gttcatccag ctcgtgcaga cgtacaacca
     3781 gctcttcgag gagaacccca ttaatgcgtc aggcgtcgac gcgaaggcta tcctgtccgc
     3841 taggctctcg aagtctcggc gcctcgagaa cctgatcgcc cagctgccgg gcgagaagaa
     3901 gaacggcctg ttcgggaatc tcattgcgct cagcctgggg ctcacgccca acttcaagtc
     3961 gaatttcgat ctcgctgagg acgccaagct gcagctctcc aaggacacat acgacgatga
     4021 cctggataac ctcctggccc agatcggcga tcagtacgcg gacctgttcc tcgctgccaa
     4081 gaatctgtcg gacgccatcc tcctgtctga tattctcagg gtgaacaccg agattacgaa
     4141 ggctccgctc tcagcctcca tgatcaagcg ctacgacgag caccatcagg atctgaccct
     4201 cctgaaggcg ctggtcaggc agcagctccc cgagaagtac aaggagatct tcttcgatca
     4261 gtcgaagaac ggctacgctg ggtacattga cggcggggcc tctcaggagg agttctacaa
     4321 gttcatcaag ccgattctgg agaagatgga cggcacggag gagctgctgg tgaagctcaa
     4381 tcgcgaggac ctcctgagga agcagcggac attcgataac ggcagcatcc cacaccagat
     4441 tcatctcggg gagctgcacg ctatcctgag gaggcaggag gacttctacc ctttcctcaa
     4501 ggataaccgc gagaagatcg agaagattct gactttcagg atcccgtact acgtcggccc
     4561 actcgctagg ggcaactccc gcttcgcttg gatgacccgc aagtcagagg agacgatcac
     4621 gccgtggaac ttcgaggagg tggtcgacaa gggcgctagc gctcagtcgt tcatcgagag
     4681 gatgacgaat ttcgacaaga acctgccaaa tgagaaggtg ctccctaagc actcgctcct
     4741 gtacgagtac ttcacagtct acaacgagct gactaaggtg aagtatgtga ccgagggcat
     4801 gaggaagccg gctttcctgt ctggggagca gaagaaggcc atcgtggacc tcctgttcaa
     4861 gaccaaccgg aaggtcacgg ttaagcagct caaggaggac tacttcaaga agattgagtg
     4921 cttcgattcg gtcgagatct ctggcgttga ggaccgcttc aacgcctccc tggggaccta
     4981 ccacgatctc ctgaagatca ttaaggataa ggacttcctg gacaacgagg agaatgagga
     5041 tatcctcgag gacattgtgc tgacactcac tctgttcgag gaccgggaga tgatcgagga
     5101 gcgcctgaag acttacgccc atctcttcga tgacaaggtc atgaagcagc tcaagaggag
     5161 gaggtacacc ggctggggga ggctgagcag gaagctcatc aacggcattc gggacaagca
     5221 gtccgggaag acgatcctcg acttcctgaa gagcgatggc ttcgcgaacc gcaatttcat
     5281 gcagctgatt cacgatgaca gcctcacatt caaggaggat atccagaagg ctcaggtgag
     5341 cggccagggg gactcgctgc acgagcatat cgcgaacctc gctggctcgc cagctatcaa
     5401 gaaggggatt ctgcagaccg tgaaggttgt ggacgagctg gtgaaggtca tgggcaggca
     5461 caagcctgag aacatcgtca ttgagatggc ccgggagaat cagaccacgc agaagggcca
     5521 gaagaactca cgcgagagga tgaagaggat cgaggagggc attaaggagc tggggtccca
     5581 gatcctcaag gagcacccgg tggagaacac gcagctgcag aatgagaagc tctacctgta
     5641 ctacctccag aatggccgcg atatgtatgt ggaccaggag ctggatatta acaggctcag
     5701 cgattacgac gtcgatcata tcgttccaca gtcattcctg aaggatgact ccattgacaa
     5761 caaggtcctc accaggtcgg acaagaaccg gggcaagtct gataatgttc cttcagagga
     5821 ggtcgttaag aagatgaaga actactggcg ccagctcctg aatgccaagc tgatcacgca
     5881 gcggaagttc gataacctca caaaggctga gaggggcggg ctctctgagc tggacaaggc
     5941 gggcttcatc aagaggcagc tggtcgagac acggcagatc actaagcacg ttgcgcagat
     6001 tctcgactca cggatgaaca ctaagtacga tgagaatgac aagctgatcc gcgaggtgaa
     6061 ggtcatcacc ctgaagtcaa agctcgtctc cgacttcagg aaggatttcc agttctacaa
     6121 ggttcgggag atcaacaatt accaccatgc ccatgacgcg tacctgaacg cggtggtcgg
     6181 cacagctctg atcaagaagt acccaaagct cgagagcgag ttcgtgtacg gggactacaa
     6241 ggtttacgat gtgaggaaga tgatcgccaa gtcggagcag gagattggca aggctaccgc
     6301 caagtacttc ttctactcta acattatgaa tttcttcaag acagagatca ctctggccaa
     6361 tggcgagatc cggaagcgcc ccctcatcga gacgaacggc gagacggggg agatcgtgtg
     6421 ggacaagggc agggatttcg cgaccgtcag gaaggttctc tccatgccac aagtgaatat
     6481 cgtcaagaag acagaggtcc agactggcgg gttctctaag gagtcaattc ggcctaagcg
     6541 gaacagcgac aagctcatcg cccgcaagaa ggactgggat ccgaagaagt acggcgggtt
     6601 cgtcagcccc actgtggcct actcggtcct ggttgtggcg aaggttgaga agggcaagtc
     6661 caagaagctc aagagcgtga aggagctgct ggggatcacg attatggagc gctccagctt
     6721 cgagaagaac ccgatcgatt tcctggaggc gaagggctac aaggaggtga agaaggacct
     6781 gatcattaag ctccccaagt actcactctt cgagctggag aacggcagga agcggatgct
     6841 ggcttccgct cgcttcctgc agaaggggaa cgagctggct ctgccgtcca agtatgtgaa
     6901 cttcctctac ctggcctccc actacgagaa gctcaagggc agccccgagg acaacgagca
     6961 gaagcagctg ttcgtcgagc agcacaagca ttacctcgac gagatcattg agcagatttc
     7021 cgagttctcc aagcgcgtga tcctggccga cgcgaatctg gataaggtcc tctccgcgta
     7081 caacaagcac cgcgacaagc caatcaggga gcaggctgag aatatcattc atctcttcac
     7141 cctgacgaac ctcggcgccc ctcgtgcttt caagtacttc gacacaacta tcgatcgcaa
     7201 ggtatacaga agcactaagg aggtcctgga cgcgaccctc atccaccagt cgattaccgg
     7261 cctctacgag acgcgcatcg acctgtctca gctcgggggc gactccggcg gcagcccaaa
     7321 gaagaagcgg aaggtgtctg gaggttctcc taagaaaaag agaaaagtgt ccggcggctc
     7381 cccgaagaag aagcgcaagg tgtaggaatt cggtacgctg aaatcaccag tctctctcta
     7441 caaatctatc tctctctatt ttctccataa ataatgtgtg agtagtttcc cgataaggga
     7501 aattagggtt cttatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt
     7561 atttgtattt gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag
     7621 tactaaaatc cagatctcct aaagtcccta tagatctttg tcgtgaatat aaaccagaca
     7681 cgagacgact aaacctggag cccagacgcc gttcgaagct agaagtaccg cttaggcagg
     7741 aggccgttag ggaaaagatg ctaaggcagg gttggttacg ttgactcccc cgtaggtttg
     7801 gtttaaatat gatgaagtgg acggaaggaa ggaggaagac aaggaaggat aaggttgcag
     7861 gccctgtgca aggtaagaag atggaaattt gatagaggta cgctactata cttatactat
     7921 acgctaaggg aatgcttgta tttataccct atacccccta ataacccctt atcaatttaa
     7981 gaaataatcc gcataagccc ccgcttaaaa attggtatca gagccatgaa taggtctatg
     8041 accaaaactc aagaggataa aacctcacca aaatacgaaa gagttcttaa ctctaaagat
     8101 aaaagatctt tcaagatcaa aactagttcc ctcacaccgg tgacggggat cgcatgcgat
     8161 atctcgagat ctagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     8221 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     8281 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     8341 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     8401 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     8461 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     8521 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     8581 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     8641 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     8701 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     8761 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     8821 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     8881 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     8941 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     9001 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     9061 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     9121 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     9181 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     9241 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     9301 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     9361 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     9421 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     9481 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     9541 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     9601 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     9661 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     9721 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     9781 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     9841 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     9901 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     9961 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    10021 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    10081 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    10141 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    10201 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    10261 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    10321 tttccccgaa aagtgccacc tgccagtgcc aagctaattc
//
